LOCUS       sequence-229824-        7330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7330)
  AUTHORS   Shpargel KB, Ospina JK, Tucker KE, Matera AG, Hebert MD
  TITLE     Control of Cajal body number is mediated by the coilin C-terminus.
  JOURNAL   J Cell Sci. 2003 Jan 15;116(Pt 2):303-12.
  PUBMED    12482916
REFERENCE   2  (bases 1 to 7330)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Sci."; date: "2003-01-15"; volume: "116(Pt 2)"; pages: "303-12"
COMMENT     Sequence Label: pEGFP-coilin
FEATURES             Location/Qualifiers
     source          1..7330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4058..4179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4095..4114)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4149..4168
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4186..4641)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4323..4344)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4535..4554
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4668..4772
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4774..5131
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4774..4965
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4797..4817)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4982..5117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5166..5960
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5166..5960
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5220..5239)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5830..5849
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6148..6167)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6192..6239
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6568..7156
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7057..7076
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt actcagatcc accaagcaag atggcagctt ccgagacggt taggctacgg
     1321 cttcaatttg attacccgcc gccagctacc ccgcactgta cggccttctg gcttctggtc
     1381 gacttgaaca gatgccgagt cgtcacagat ctcattagtc tcatccgcca gcgcttcggc
     1441 ttcagttctg gggccttcct aggcctctac ctggaggggg ggctcttgcc ccccgccgag
     1501 agcgcgcgcc ttgtgagaga caacgactgc ctcagagtta aattagaaga gagaggagtt
     1561 gctgagaatt ctgtagtcat cagtaatggt gacattaatt tatctcttag aaaagcaaag
     1621 aagcgggcat ttcagttaga ggagggtgaa gaaactgaac cagattgcaa atattcaaag
     1681 aagcattgga agagtcgaga gaacaataac aataatgaga aggtcttgga tctggaacca
     1741 aaagctgtca cagatcagac tgtcagcaaa aaaaacaaga gaaaaaataa agcaacctgt
     1801 ggcacagtgg gtgatgataa cgaagaggcc aaaagaaaat caccaaagaa aaaggagaaa
     1861 tgtgaatata aaaaaaaggc taagaatccc aagtctccga aagtacaggc agtgaaagac
     1921 tgggccaatc agagatgtag ttctccaaaa ggttctgcta gaaacagcct tgttaaagcc
     1981 aaaaggaaag gtagtgtaag cgtttgctca aaagagagtc ccagttcctc ctcggagtct
     2041 gagtcttgtg atgaatctat cagtgatggt cccagcaaag tcactttgga ggccagaaat
     2101 tcctcagaga aattaccaac tgagttatca aaggaagaac cctctaccaa aaatacaact
     2161 gcagacaaac tggctataaa acttggcttt agccttaccc ccagcaaggg caagacctct
     2221 ggaacaacat cttccagttc agactctagt gcagagtcag acgaccaatg cttgatgtca
     2281 tcgagcaccc cggagtgtgc tgcgggtttc ttaaagacag taggcctttt tgcaggaaga
     2341 ggtcgtccag gcccagggct gtcatcacag actgcaggtg ctgctggatg gaggcgttct
     2401 ggctcaaatg gtggtggaca ggctcctggt gcttctccca gtgtgtctct ccctgctagt
     2461 ttaggaagag gatggggtag agaagagaac cttttttctt ggaagggagc taagggacgg
     2521 ggcatgcggg ggagaggtcg aggacgaggg catcctgttt cctgtgttgt aaatagaagc
     2581 actgacaacc agaggcaaca gcaattaaat gacgtggtaa aaaattcatc tactattatc
     2641 cagaatccag tagagacacc caagaaggac tatagtctgt taccactgtt agcagctgcc
     2701 cctcaagttg gagaaaagat tgcatttaag cttttggagc taacatccag ttactctcct
     2761 gatgtctctg actacgagga aggaagaata ttaagccaca atccagagac ccagcaagta
     2821 gatatagaaa ttctttcatc cttacctgcc ttgagagaac ctgggaaatt tgatttagtt
     2881 tatcacaatg aaaatggagc tgaggtagtg gagtacgctg tgacacagga gagcaagatc
     2941 actgtatttt ggaaagagtt gattgaccca agactgatta ttgaatctcc aagtaacaca
     3001 tcaagtacag aacctgcctg agtatgacct ctccacctca tagtttatga atgtcttgtt
     3061 tgtgaaagtg actataaccc aaactttttt tttttttaaa gaggatttgg aagttgtatg
     3121 gatttttttg ttatcttcac tttactgcat aggaaacaat ctacctcatc atttaaaatg
     3181 acatgggtgt cggttttgta gatctttggt ttttttgtca ggtttaattt cagttaacaa
     3241 aatgtaaaac atgacattcc ctgcagatat tgttgtatgc cagtatggtt tcttctcttt
     3301 ctttaaatgt ttttggccat caagtagcag tcgtcagtag gagtttataa taccaagaat
     3361 gtgctgcgta tcttgtctca ataagtttta agtaacattt aaaaatatta aagcatgtta
     3421 tttgacctaa ttttttagca tttgagttgt tccattaaat ggagcatctt gtaaatttca
     3481 agtattttat acttgcaatt gttaagagtt aacaggtagt tggatttgtc gcagacaatg
     3541 agttaaggaa tcctttcacg tttttcccaa ctttaaaatt aaggattctc aggaccctgt
     3601 gtagagcagt gaaaataaga tgtgcgtatg tgtgtgtatg cctggagaat tggtgtttca
     3661 cttcagtgag aggattggct gtgagcttca gaccaggaaa tgtgtcatct tgccaggcac
     3721 ctggctgagt gtgctggagt gaggatcttg aacagaaact tccttttctg ttattattca
     3781 ctacgaagct aaaatggcca aatatatacc gtgaaaattg gtttcattta acaaaagatc
     3841 agatccctcc ttcagctgta tctagagcgg ccgccaccgc ggtggagctc aagcttcgaa
     3901 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat
     3961 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4021 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4081 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4141 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     4201 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4261 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4321 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4381 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4441 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4501 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4561 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4621 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4681 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4741 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4801 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4861 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4921 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4981 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5041 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5101 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     5161 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5221 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5281 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5341 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5401 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5461 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5521 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5581 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5641 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     5701 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5761 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5821 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5881 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5941 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6001 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6061 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6121 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     6181 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     6241 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     6301 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     6361 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     6421 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6481 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6541 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6601 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6661 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6721 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6781 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6841 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6901 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6961 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7021 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7081 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7141 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7201 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7261 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     7321 tggagttccg
//
